LOCUS       sequence-12370-e        9539 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9539)
  TITLE     New vectors Aug. 2010
REFERENCE   2  (bases 1 to 9539)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pENTR-GFP-MDC1 (770-7)
FEATURES             Location/Qualifiers
     source          1..9539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      498..507
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(504..506,507..509,510..1220)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(504..506,507..509,510..1220)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(549..570)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(810..829)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1157..1178
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(7768..7867)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7979..7998)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             7990..8799
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(8057..8076)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      8889..9477
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9378..9397
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggctcca ccatgggaac caattcagtc
      481 gactagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
      541 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
      601 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
      661 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
      721 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
      781 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
      841 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
      901 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
      961 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     1021 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1081 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     1141 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1201 gcatggacga gctgtacaag tccggccgga ctcagatctc gagctcaagc ttcgaattct
     1261 gcagtcgacg gtaccgcggg cccatcatgg aggacaccca ggctattgac tgggatgttg
     1321 aagaagagga ggagacagag caatccagtg aatccttgag gtgtaacgtg gagccagtag
     1381 ggcggctaca tatctttagt ggtgcccatg gaccagaaaa agatttccca ctacacctcg
     1441 ggaagaatgt ggtaggccga atgcctgact gctctgtggc cctgcccttt ccatctatct
     1501 ccaaacaaca tgcagagatt gaaatcttag cctgggacaa ggcacctatc ctccgagact
     1561 gtgggagcct taatggtact caaatcctga gacctcctaa ggttttgagc cctggggtga
     1621 gtcaccgtct gagggaccag gaattgattc tctttgctga cttgctctgc cagtaccatc
     1681 gcctggatgt ctctctgccc tttgtctccc ggggccctct gacagtagaa gagacaccca
     1741 gagtacaggg agaaactcaa ccccagaggc ttctgttggc tgaggactcg gaggaggaag
     1801 tagattttct ttctgaaagg cgtatggtaa aaaaatcaag gaccacatct tcctctgtga
     1861 tagttccaga gagtgatgaa gaggggcatt ccccggtcct gggcggcctt gggccgcctt
     1921 ttgccttcaa tttgaacagt gacacagatg tggaagaagg tcagcaacca gccacagagg
     1981 aggcctcctc agctgccaga agaggtgcca ctgtagaggc aaagcagtct gaagctgaag
     2041 ttgtaactga aatccagctt gaaaaggatc agcctttagt gaaggagagg gacaatgata
     2101 caaaagtcaa gaggggtgca gggaatgggg tggttccagc tggggtgatt ctggagagga
     2161 gccaacctcc tggagaggac agtgacacag atgtggatga tgacagcagg cctcctggaa
     2221 ggccagctga ggtccatttg gaaagggctc agccttttgg cttcatcgac agcgacactg
     2281 atgcggaaga agagaggatc ccagcaaccc cagttgtcat tcctatgaag aagaggaaga
     2341 tcttccatgg agtaggtaca aggggtcctg gagcaccagg cctggcccat ctgcaggaga
     2401 gccaggctgg tagtgataca gatgtggaag aaggcaaggc cccacaggct gtccctctgg
     2461 agaaaagcca agcttccatg gttatcaaca gcgatacaga tgacgaggaa gaagtctcag
     2521 cagcgctgac tttggcacat ctgaaagaga gccagcctgc tatatggaac agagatgcag
     2581 aagaggacat gccccaacgt gtggtccttc tgcagcgaag ccaaaccacc actgagagag
     2641 acagtgacac agacgtggag gaggaagagc tcccagtgga aaatagagaa gctgtcctca
     2701 aggatcacac aaagattaga gcccttgtta gagcacattc agaaaaggac caacctcctt
     2761 ttggggacag tgatgacagt gtggaagcag ataagagctc acctgggatc cacctggaga
     2821 gaagccaagc ctccaccaca gtggacatca acacacaagt ggagaaggaa gtcccgccag
     2881 ggtcagccat tatgcatata aagaagcatc aggtgtctgt ggaggggaca aatcaaacag
     2941 atgtgaaagc agttggggga ccagcaaagc tgcttgtggt atctctagag gaagcctggc
     3001 ctctgcatgg ggactgtgaa acagatgcag aggagggcac ctccctaaca gcctcagtag
     3061 ttgcagatgt aagaaagagc cagcttccag cagaagggga tgctggggca gagtgggctg
     3121 cagctgttct taagcaggag agagctcatg aggtgggggc ccagggtggg ccacctgtgg
     3181 cacaagtgga gcaggacctc cctatctcaa gagagaacct cacagatctg gtggtggaca
     3241 cagacactct aggggaatcc acccagccac agagagaggg agcccaggtc cccacaggaa
     3301 gggagagaga acaacatgtg ggtgggacca aggactctga agacaactat ggtgattctg
     3361 aagatctgga cctacaagct acccagtgct ttctggagaa tcagggcctg gaagcagtcc
     3421 agagcatgga ggatgaacct acccaggcct tcatgttgac tccaccccaa gagcttggcc
     3481 cttcccattg cagcttccag acaacaggta ccctagatga accatgggag gtcctggcta
     3541 cacagccatt ctgtctgaga gagtctgagg actctgagac ccagcctttt gacacgcacc
     3601 ttgaggccta tggaccttgc ctgtctccac ctagggcaat accaggagac caacatccag
     3661 agagcccagt tcacacagag ccaatgggga ttcaaggcag agggaggcag actgtggata
     3721 aagtcatggg tataccaaaa gaaacagcag agagggtggg ccctgagaga gggccattgg
     3781 agagagaaac tgagaaactg ctaccagaaa gacagacaga tgtgacagga gaggaagaat
     3841 taaccaaggg gaaacaggac agagaacaaa aacagttgtt agctagagac acccagagac
     3901 aagaatctga caaaaatggg gaaagtgcaa gtcctgaaag agatagggag agtttgaagg
     3961 tagaaattga gacatctgag gaaatacaag agaaacaagt acagaagcag acccttccaa
     4021 gcaaagcatt tgagagagaa gtagagagac cagtagcaaa cagagagtgc gatccagccg
     4081 agttagaaga gaaggtgccc aaagtgatcc tggagagaga tacacagaga ggggagccag
     4141 agggagggag ccaggaccag aaagggcagg cctccagccc aacaccagag cctggggtgg
     4201 gggcggggga ccttccggga cctacctcag cccccgtacc ttctgggagc cagtcaggtg
     4261 gaaggggatc cccagtgagc cccaggaggc atcagaaagg cctcctgaat tgcaagatgc
     4321 cacctgctga gaaggcttcc aggatcagag ctgctgagaa ggtttccagg ggcgatcagg
     4381 aatctccaga tgcttgtctg cctcctgcag tacctgaagc cccagcccca ccccaaaagc
     4441 cccttaactc tcagagccag aaacatcttg cacctccgcc ccttctttct ccccttttac
     4501 cttctatcaa gccaaccgtt cgtaagacca ggcaagatgg gagtcaggaa gctccagagg
     4561 ctcccttgtc ctcagagctg gagcctttcc acccaaagcc taaaattaga actcggaagt
     4621 cctccagaat gacacccttt ccagctacct ctgctgcccc tgagccccac ccttccacct
     4681 ccacagccca gccagtcact cccaagccca catctcaggc cactaggagc aggacaaata
     4741 ggtcctctgt caagacccct gaaccagttg tccccacagc ccctgagctc cagccttcca
     4801 cctccacaga ccagcctgtc acctctgagc ccacatctca ggttactagg ggaagaaaaa
     4861 gtagatcctc tgtcaagacc cctgaaacag ttgtgcccac agcccttgag ctccagcctt
     4921 ccacctccac cgaccgacct gtcacctctg aacccacctc tcaggctact aggggaagaa
     4981 aaaatagatc ctctgtcaag acccctgaac cagttgtccc cacagcccct gagctccagc
     5041 cttccacctc cacagaccag cctgtcactt ctgagcccac atatcaggct actaggggaa
     5101 gaaaaaatag atcctctgtc aagacccctg aaccagttgt gcccacagcc cctgagctcc
     5161 ggccttccac ctccacagac cgacctgtca cccccaagcc cacatctcgg accactagga
     5221 gcaggacaaa tatgtcctct gtcaagaccc ctgaaacagt tgtccccaca gcccctgagc
     5281 tccagatttc cacctccaca gaccaacctg tcacccctaa gcccacatct cggaccacta
     5341 ggagcaggac aaatatgtcc tctgtgaaga accctgaatc aactgtccct atagcccctg
     5401 agctcccacc ttccacctcc acagagcagc ctgtcacccc tgagcccaca tctcgggcta
     5461 ctaggggaag aaaaaataga tcctctggca agacccctga aacacttgtc cccacagccc
     5521 ctaagctcga gccttccact tccacagacc aacctgtcac tcctgagccc acatctcagg
     5581 ccaccagggg caggacaaat aggtcctctg tgaagacccc tgaaacagtt gtccccacag
     5641 cccctgagct ccagccttcc acctccacag accagcctgt tacccctgag cctacgtctc
     5701 aggctactag gggaagaaca gatagatcct ctgtcaagac tcctgaaaca gttgtcccca
     5761 cagcccctga gctacaggct tccgcctcca cagaccagcc tgtcacctct gagcccacat
     5821 ctcggaccac taggggaaga aaaaatcggt cctctgtcaa gacccctgaa acagttgtgc
     5881 ccgcagcccc tgagctccag cctcccacct ccacagaccg acctgtcacc cctgagccca
     5941 catctcgggc cactaggggc aggacaaata ggtcctctgt caagacccct gaatcaattg
     6001 tccctatagc ccctgagctt cagccttcca cctccagaaa ccagcttgtc acccctgagc
     6061 ccacatctcg ggccactagg tgcaggacaa ataggtcctc tgtcaagacc cctgagccag
     6121 ttgtccccac agcccctgag ccccatccta ccacctccac agaccagcct gtcaccccca
     6181 agctcacatc tagggccact aggagaaaga caaataggtc ctctgtcaag actcccaaac
     6241 cagttgaacc agcagcctct gatcttgagc cttttacccc cacagaccag tccgtcaccc
     6301 ctgaggccat agctcagggt ggtcagagca aaacactgag gtcttccaca gtaagagcta
     6361 tgccggttcc taccacccct gaattccaat ctcctgtcac cacagaccag cctatttccc
     6421 ctgagcctat tactcaaccc agttgcatca agaggcagag agccgctggg aaccctggct
     6481 ccctcgcagc tcccattgac cataagcctt gctctgcacc cttggaacct aaatcccagg
     6541 cctcaaggaa ccaaagatgg ggagcagtga gagcagctga atcccttaca gccattcctg
     6601 agcctgcctc tccccagctt cttgagacac caattcatgc ctcccagatc caaaaggtgg
     6661 aaccagcagg tagatctagg ttcaccccgg agctccagcc taaggcctct caaagccgca
     6721 agaggtcttt agctaccatg gattcaccac cacatcaaaa acagccccaa agaggggaag
     6781 tctcccagaa gacagtgatt atcaaggaag aggaagaaga tactgcagag aagccaggga
     6841 aggaagagga tgtcgtgact ccaaaaccag gcaagagaaa gagagaccag gcagaggagg
     6901 agcccaacag aataccaagc cgcagcctcc gacggaccaa acttaaccaa gaatcaacag
     6961 cccccaaagt gctcttcaca ggagtggtgg atgctcgggg agagcgggct gtgctggcac
     7021 tggggggaag tctggctggt tcagcggcag aggcttccca cctggtcact gatcgcatcc
     7081 gccggacagt caagttcctg tgtgccctgg ggcggggaat ccccattctg tccctggact
     7141 ggctgcatca gtcccgcaag gctggtttct tcttaccccc ggatgaatat gtggtgaccg
     7201 accctgagca agagaagaac tttggcttta gccttcaaga cgcactgagc agggctcggg
     7261 agcgaaggct gctagagggc tatgagatct atgtgacccc tggagtccag ccaccaccac
     7321 ctcagatggg agagattatt agctgctgtg gaggcacata cctacccagc atgcctcggt
     7381 cctataagcc tcagagagtt gtgatcacat gccctcagga cttccctcat tgctccattc
     7441 cactacgggt tgggctgccc ctcctctcgc ctgagttcct gctgactgga gtgctgaagc
     7501 aggaagccaa gccagaggcc tttgtcctct cccctttgga gatgtcatcc acctgagaac
     7561 tccactaccc ttttccctcc cagaccacga attagaagat atgtggaaga aagaactcag
     7621 ggcgttagaa aggattgggg tatattgata caacttgtcc tggaacatgg gtgggaccag
     7681 aaatctttat gaataaatga aaagataagg gatttggaag ccacaggttg ttttttgttt
     7741 gtttgtttgt ttttttaatg gatctagacc cagctttctt gtacaaagtt ggcattataa
     7801 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca
     7861 ttatttgcca tccagctgca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     7921 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     7981 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     8041 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     8101 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     8161 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     8221 cctcttccga ccatcaagca ttttatccgt actcctggtg atgcatggtt actcaccact
     8281 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     8341 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     8401 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     8461 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     8521 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     8581 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     8641 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     8701 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     8761 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     8821 taacattatt cagattgggc cccgttccac tgagcgtcag accccgtaga aaagatcaaa
     8881 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     8941 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     9001 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     9061 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     9121 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     9181 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     9241 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     9301 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     9361 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     9421 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     9481 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
//
